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Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Leite C, Silva NT, Mendes S, Ribeiro A, de Faria JP, Lourenço T, dos Santos F, Andrade PZ, Cardoso CM, Vieira M, Paiva A, da Silva CL, Cabral JM, Relvas JB, Grãos M - PLoS ONE (2014)

Bottom Line: In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples.The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity.The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

View Article: PubMed Central - PubMed

Affiliation: Biocant - Technology Transfer Association, Biocant Park, Cantanhede, Portugal.

ABSTRACT
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

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Immunofluorescence microscopy images of OL-like cells derived from hUCM-MSCs in co-culture with mouse dorsal root ganglion (DRG) neurons.The co-cultures were stained using an anti-human Golgi antibody (green) to specifically identify the human cells (OL-like cells derived form MSCs) and an anti-beta-III-tubulin antibody (red) to label the mouse DRG neurons. The images acquired in the green channel (anti-human Golgi antibody) were deliberately slightly overexposed to allow for a better understanding of the cellular morphology, which did not affect the identification of human versus mouse cells, as evidenced by the lack of green signal in the mouse DRG neurons (in red). Counterstaining of the nuclei was performed using DAPI (blue). It was apparent that the OL-like cells and DRG neurons tend to cluster together, as illustrated in a lower magnification image (A). Higher magnification images (B-D) suggest the existence of contact points (arrows) between branches of OL-like cells displaying an immature oligodendrocyte-like morphology (asterisks) and neurites. Scale bars correspond to 50 µm (A) or 20 µm (B-D).
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pone-0111059-g009: Immunofluorescence microscopy images of OL-like cells derived from hUCM-MSCs in co-culture with mouse dorsal root ganglion (DRG) neurons.The co-cultures were stained using an anti-human Golgi antibody (green) to specifically identify the human cells (OL-like cells derived form MSCs) and an anti-beta-III-tubulin antibody (red) to label the mouse DRG neurons. The images acquired in the green channel (anti-human Golgi antibody) were deliberately slightly overexposed to allow for a better understanding of the cellular morphology, which did not affect the identification of human versus mouse cells, as evidenced by the lack of green signal in the mouse DRG neurons (in red). Counterstaining of the nuclei was performed using DAPI (blue). It was apparent that the OL-like cells and DRG neurons tend to cluster together, as illustrated in a lower magnification image (A). Higher magnification images (B-D) suggest the existence of contact points (arrows) between branches of OL-like cells displaying an immature oligodendrocyte-like morphology (asterisks) and neurites. Scale bars correspond to 50 µm (A) or 20 µm (B-D).

Mentions: By the end of the co-culture period, it could be observed that OL-like cells typically tended to be present in the vicinity of neurons (Figure 9A - neurons stained in red for beta-III-tubulin and OL-like cells stained in green for human Golgi). This observation suggests that there might occur a positive crosstalk between both types of cells (neurons and OL-like cells).


Differentiation of human umbilical cord matrix mesenchymal stem cells into neural-like progenitor cells and maturation into an oligodendroglial-like lineage.

Leite C, Silva NT, Mendes S, Ribeiro A, de Faria JP, Lourenço T, dos Santos F, Andrade PZ, Cardoso CM, Vieira M, Paiva A, da Silva CL, Cabral JM, Relvas JB, Grãos M - PLoS ONE (2014)

Immunofluorescence microscopy images of OL-like cells derived from hUCM-MSCs in co-culture with mouse dorsal root ganglion (DRG) neurons.The co-cultures were stained using an anti-human Golgi antibody (green) to specifically identify the human cells (OL-like cells derived form MSCs) and an anti-beta-III-tubulin antibody (red) to label the mouse DRG neurons. The images acquired in the green channel (anti-human Golgi antibody) were deliberately slightly overexposed to allow for a better understanding of the cellular morphology, which did not affect the identification of human versus mouse cells, as evidenced by the lack of green signal in the mouse DRG neurons (in red). Counterstaining of the nuclei was performed using DAPI (blue). It was apparent that the OL-like cells and DRG neurons tend to cluster together, as illustrated in a lower magnification image (A). Higher magnification images (B-D) suggest the existence of contact points (arrows) between branches of OL-like cells displaying an immature oligodendrocyte-like morphology (asterisks) and neurites. Scale bars correspond to 50 µm (A) or 20 µm (B-D).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214693&req=5

pone-0111059-g009: Immunofluorescence microscopy images of OL-like cells derived from hUCM-MSCs in co-culture with mouse dorsal root ganglion (DRG) neurons.The co-cultures were stained using an anti-human Golgi antibody (green) to specifically identify the human cells (OL-like cells derived form MSCs) and an anti-beta-III-tubulin antibody (red) to label the mouse DRG neurons. The images acquired in the green channel (anti-human Golgi antibody) were deliberately slightly overexposed to allow for a better understanding of the cellular morphology, which did not affect the identification of human versus mouse cells, as evidenced by the lack of green signal in the mouse DRG neurons (in red). Counterstaining of the nuclei was performed using DAPI (blue). It was apparent that the OL-like cells and DRG neurons tend to cluster together, as illustrated in a lower magnification image (A). Higher magnification images (B-D) suggest the existence of contact points (arrows) between branches of OL-like cells displaying an immature oligodendrocyte-like morphology (asterisks) and neurites. Scale bars correspond to 50 µm (A) or 20 µm (B-D).
Mentions: By the end of the co-culture period, it could be observed that OL-like cells typically tended to be present in the vicinity of neurons (Figure 9A - neurons stained in red for beta-III-tubulin and OL-like cells stained in green for human Golgi). This observation suggests that there might occur a positive crosstalk between both types of cells (neurons and OL-like cells).

Bottom Line: In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples.The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity.The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

View Article: PubMed Central - PubMed

Affiliation: Biocant - Technology Transfer Association, Biocant Park, Cantanhede, Portugal.

ABSTRACT
Mesenchymal stem cells (MSCs) are viewed as safe, readily available and promising adult stem cells, which are currently used in several clinical trials. Additionally, their soluble-factor secretion and multi-lineage differentiation capacities place MSCs in the forefront of stem cell types with expected near-future clinical applications. In the present work MSCs were isolated from the umbilical cord matrix (Wharton's jelly) of human umbilical cord samples. The cells were thoroughly characterized and confirmed as bona-fide MSCs, presenting in vitro low generation time, high proliferative and colony-forming unit-fibroblast (CFU-F) capacity, typical MSC immunophenotype and osteogenic, chondrogenic and adipogenic differentiation capacity. The cells were additionally subjected to an oligodendroglial-oriented step-wise differentiation protocol in order to test their neural- and oligodendroglial-like differentiation capacity. The results confirmed the neural-like plasticity of MSCs, and suggested that the cells presented an oligodendroglial-like phenotype throughout the differentiation protocol, in several aspects sharing characteristics common to those of bona-fide oligodendrocyte precursor cells and differentiated oligodendrocytes.

Show MeSH
Related in: MedlinePlus